Assembly and infection efficacy of hepatitis B virus surface protein exchanges in 8 hepatitis D virus genotype isolates
Assembly and infection efficacy of hepatitis B virus surface protein exchanges in 8 hepatitis D virus genotype isolates
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8株丁型肝炎病毒基因型分离株乙型肝炎病毒表面蛋白交换的组装及感染效果
DOI：
10.1016/j.jhep.2021.03.025
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发表时间：
2021-07-16
影响因子：
25.7
通讯作者：
Urban, Stephan
中科院分区：
文献类型：
--
作者：
Wang, Wenshi;Lempp, Florian A.;Urban, Stephan
背景与目的：慢性HDV感染是最严重的病毒性肝炎。HDV需要HBV膜蛋白进入肝细胞、组装颗粒和释放。目前已鉴定出8种HDV和8种HBV型。方法：我们亚克隆了HDV和HBV型的互补DNA，并用Northern印迹、Western印迹、逆转录定量聚合酶链式反应和细胞内ELISA对HDV的复制、组装和感染性进行了系统的研究。结果：8个HDV克隆启动了HDV的复制，复制效率有显著差异。这8个HBVHBs抗原编码结构都支持亚病毒颗粒的分泌，但观察到包膜蛋白化学计量和分泌效率的差异。所有HDV/HBV组合的共转染支持颗粒组装，然而，各自的伪型HDV在组装动力学方面有所不同。最高产的组合与自然地理分布无关，反对HDV核糖核蛋白复合体对乙肝病毒包膜的进化适应。所有的HDV都能引起强大的、可比的先天免疫反应。观察到EMA批准的进入抑制剂Buevitide的活性依赖于HBV膜的差异，然而，在治疗剂量下可以实现有效的抑制。结论：不同基因型的HDV复制效果不同。HDV基因组的变异和HBV膜蛋白的变异都是HDV出入境效应的主要决定因素，从而决定了Lonafarnib对HDV的组装抑制或Buevitide对HDV进入的抑制。这些差异可能影响HDV的致病性、免疫应答和新的药物治疗方案的疗效。我们研究了不同HDV基因型别感染细胞和复制的能力。我们还评估了来自不同HBV型的包膜蛋白对HDV感染性和复制的影响。在此，我们证实了HDV和HBV膜蛋白的基因差异是HDV组装、从头进入细胞以及新型抗病毒药物疗效的主要决定因素。(C)2021年提交人(S)。由Elsevier B.V.代表欧洲肝脏研究协会出版。
Background & Aims: Chronic HDV infections cause the most severe form of viral hepatitis. HDV requires HBV envelope proteins for hepatocyte entry, particle assembly and release. Eight HDV and 8 HBV genotypes have been identified. However, there are limited data on the replication competence of different genotypes and the effect that different HBV envelopes have on virion assembly and infectivity.Methods: We subcloned complementary DNAs (cDNAs) of all HDV and HBV genotypes and systematically studied HDV replication, assembly and infectivity using northern blot, western blot, reverse-transcription quantitative PCR, and in-cell ELISA.Results: The 8 HDV cDNA clones initiated HDV replication with noticeable differences regarding replication efficacy. The 8 HBVHBsAg-encoding constructs all supported secretion of subviral particles, however variations in envelope protein stoichiometry and secretion efficacy were observed. Co-transfection of all HDV/ HBV combinations supported particle assembly, however, the respective pseudo-typed HDVs differed with respect to assembly kinetics. The most productive combinations did not correlate with the natural geographic distribution, arguing against an evolutionary adaptation of HDV ribonucleoprotein complexes to HBV envelopes. All HDVs elicited robust and comparable innate immune responses. HBV envelope-dependent differences in the activity of the EMA-approved entry inhibitor bulevirtide were observed, however efficient inhibition could be achieved at therapeutically applied doses. Lonafarnib also showed pan-genotypic activity.Conclusions: HDVs from different genotypes replicate with variable efficacies. Variations in HDV genomes and HBV envelope proteins are both major determinants of HDV egress and entry efficacy, and consequently assembly inhibition by lonafarnib or entry inhibition by bulevirtide. These differences possibly influ-ence HDV pathogenicity, immune responses and the efficacy of novel drug regimens.Lay summary: HDV requires the envelope protein of HBV for assembly and to infect human cells. We investigated the ability of different HDV genotypes to infect cells and replicate. We also assessed the effect that envelope proteins from different HBV genotypes had on HDV infectivity and replication. Herein, we confirmed that genotypic differences in HDV and HBV envelope proteins are major determinants of HDV assembly, de novo cell entry and consequently the efficacy of novel antivirals. (C) 2021 The Author(s). Published by Elsevier B.V. on behalf of European Association for the Study of the Liver.